U.S. patent application number 16/614152 was filed with the patent office on 2020-05-14 for inducing phospholipidosis for enhancing therapeutic efficacy.
The applicant listed for this patent is Nextcea Inc.. Invention is credited to Frank Hsieh.
Application Number | 20200147070 16/614152 |
Document ID | / |
Family ID | 64274832 |
Filed Date | 2020-05-14 |
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United States Patent
Application |
20200147070 |
Kind Code |
A1 |
Hsieh; Frank |
May 14, 2020 |
INDUCING PHOSPHOLIPIDOSIS FOR ENHANCING THERAPEUTIC EFFICACY
Abstract
A method of enhancing the efficacy of a therapeutic compound,
the method comprising administering an effective amount of a
phospholipidosis (PL)-inducing compound (or compounds) to a patient
in need of a therapeutic compound for a disorder, whereby PL is
induced in the patient; and administering the therapeutic compound
to the patient.
Inventors: |
Hsieh; Frank; (Woburn,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Nextcea Inc. |
Woburn |
MA |
US |
|
|
Family ID: |
64274832 |
Appl. No.: |
16/614152 |
Filed: |
May 16, 2018 |
PCT Filed: |
May 16, 2018 |
PCT NO: |
PCT/US2018/032885 |
371 Date: |
November 15, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62507537 |
May 17, 2017 |
|
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|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N 33/48 20130101;
A61K 31/343 20130101; G01N 33/92 20130101; A61K 9/2806 20130101;
A61K 31/138 20130101; A61K 31/47 20130101 |
International
Class: |
A61K 31/47 20060101
A61K031/47; G01N 33/92 20060101 G01N033/92; A61K 31/343 20060101
A61K031/343; A61K 31/138 20060101 A61K031/138 |
Claims
1. A method of enhancing the efficacy of a therapeutic compound,
the method comprising: administering an effective amount of a
phospholipidosis (PL)-inducing compound to a patient in need of a
therapeutic compound for a disorder, whereby PL is induced in the
patient; and administering the therapeutic compound to the
patient.
2. The method of claim 1, wherein the PL-inducing compound induces
PL in a tissue, organ, or cell affected by the disorder or intended
to be targeted by the therapeutic compound.
3. The method of claim 1, wherein the PL-inducing compound induces
PL in a tissue, organ, or cell not affected by the disorder or not
intended to be targeted by the therapeutic compound.
4. The method of claim 1, wherein the PL-inducing compound induces
multi-organ PL.
5. The method of claim 1, wherein administration of the PL-inducing
compound decreases lysosomal degradation of the therapeutic
compound.
6. The method of claim 2, wherein administration of the PL-inducing
compound increases the concentration of the therapeutic compound in
the tissue, organ, or cell.
7. The method of claim 3, wherein administration of the PL-inducing
compound reduces concentration of the therapeutic compound in the
tissue, organ, or cell.
8. The method of claim 1, wherein the PL-inducing compound and the
therapeutic compound are administered at the same time or about the
same time before PL is induced in the patient.
9. The method of claim 1, wherein the therapeutic compound is
administered only after PL is induced in the patient.
10. The method of claim 1, further comprising monitoring the
occurrence, progress, or reversibility of PL in the patient.
11. The method of claim 10, wherein the monitoring step includes
detecting the levels of one or more biomarkers in a biological
sample obtained from the patient, the one or more biomarkers being
selected from the group consisting of 2,2' di-22:6-BMP, 3,2'
di-22:6-BMP, 2,3' di-22:6-BMP, 3,3' di-22:6-BMP, di-18:1-BMP,
di-18:2-BMP, 18:1/18:2-BMP, 18:1/22:6-BMP, 18:2/22:6-BMP,
di-22:6-PG, di-18:1-PG, di-18:2-PG, 18:1/18:2-PG, 18:1/22:6-PG,
18:2/22:6-PG, mono-22:6-BMP, mono-18:1-BMP, and mono-18:2-BMP.
12. The method of claim 11, wherein the monitoring step further
includes detecting the levels of one or more additional species of
BMP, PG, or mono-BMP, or total BMP.
13. The method of claim 11, wherein the therapeutic compound is
administered after an elevated level of the biomarker, as compared
to a control level, is detected in the biological sample.
14. The method of claim 1, wherein the patient is administered with
the therapeutic compound for a treatment period and PL is induced
for a portion or over the entire duration of the treatment
period.
15. The method of claim 1, wherein the therapeutic compound is a
biological drug.
16. The method of claim 1, wherein the therapeutic compound is a
small molecule drug.
17. A pharmaceutical composition, comprising a PL-inducing compound
and a therapeutic compound for a disorder.
18. The composition of claim 17, wherein the composition is a
controlled-released formulation.
19. The composition of claim 18, wherein the formulation is
formulated to release the PL-inducing compound and the therapeutic
compound simultaneously or at different rates or times.
20. The composition of claim 19, wherein the PL-inducing compound
is released before the therapeutic compound.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims priority to U.S. Provisional
Application No. 62/507,537, filed on May 17, 2017, the entire
content of which is hereby incorporated by reference herein.
BACKGROUND
[0002] Phospholipidosis (PL) is a lysosomal storage condition
characterized by the accumulation of multi-lamellar (myeloid)
bodies in cells and tissues. It can be induced by various natural
and synthetic compounds, and is a common finding in animals and
humans treated with cationic amphiphilic drugs (CADs). PL is
typically reversible after the cessation of drug treatment. It can
be induced at a manageable level without serious collateral drug
side effects.
SUMMARY
[0003] This invention is based, at least in part, on the unexpected
discovery that PL can be used to selectively inhibit lysosomal
degradation and increase drug exposure in target tissues, cells, or
organs to enhance in vivo therapeutic efficacy.
[0004] In one aspect, a method is described for enhancing the
efficacy of a therapeutic compound. The method includes
administering an effective amount of a PL-inducing compound to a
patient in need of a therapeutic compound for a disorder, whereby
PL is induced in the patient; and administering the therapeutic
compound to the patient.
[0005] For example, the therapeutic compound can be for treating
colon cancer, breast cancer, prostate cancer, hepatocellular
carcinoma, melanoma, lung cancer, glioblastoma, brain tumor,
hematopoeitic malignancies, retinoblastoma, renal cell carcinoma,
head and neck cancer, cervical cancer, pancreatic cancer,
esophageal cancer, squama cell carcinoma, hemophila,
hypercholesterolemia, inflammatory dermatoses (e.g., dermatitis,
eczema, atopic dermatitis, allergic contact dermatitis, urticaria,
necrotizing vasculitis, cutaneous vasculitis, hypersensitivity
vasculitis, eosinophilic myositis, polymyositis, dermatomyositis,
and eosinophilic fasciitis), inflammatory bowel diseases (e.g.,
Crohn's disease and ulcerative colitis), acute respiratory distress
syndrome, fulminant hepatitis, hypersensitivity lung diseases
(e.g., hypersensitivity pneumonitis, eosinophilic pneumonia,
delayed-type hypersensitivity, interstitial lung disease or ILD,
idiopathic pulmonary fibrosis, and ILD associated with rheumatoid
arthritis), asthma, and allergic rhinitis, autoimmune diseases
(e.g., rheumatoid arthritis, psoriatic arthritis, systemic lupus
erythematosus, myasthenia gravis, juvenile onset diabetes,
glomerulonephritis, autoimmune throiditis, ankylosing spondylitis,
systemic sclerosis, and multiple sclerosis), acute and chronic
inflammatory diseases (e.g., systemic anaphylaxia or
hypersensitivity responses, drug allergies, insect sting allergies,
allograft rejection, and graft-versus-host disease), Sjogren's
syndrome, human immunodeficiency virus infection, tumor metastasis,
bronchitis, cystic fibrosis, chronic obstructive lung disease,
kidney diseases (e.g., kidney cancer, nephritis, and nephropathy),
and neurological diseases (e.g., Alzheimer's disease, Parkinson's
disease, Huntington's disease, amyotrophic lateral sclerosis).
[0006] In some embodiments, the PL-inducing compound induces PL in
a tissue, organ, or cell affected by the disorder or intended to be
targeted by the therapeutic compound. In other embodiments, the
PL-inducing compound induces PL in a tissue, organ, or cell not
affected by the disorder or not intended to be targeted by the
therapeutic compound. In some embodiments, the PL-inducing compound
induces multi-organ PL.
[0007] In some embodiments, administration of the PL-inducing
compound decreases lysosomal degradation of the therapeutic
compound. In some embodiments, administration of the PL-inducing
compound increases the concentration of the therapeutic compound in
a tissue, organ, or cell affected by the disorder or intended to be
targeted by the therapeutic compound. In some embodiments,
administration of the PL-inducing compound reduces the
concentration of the therapeutic compound in a tissue, organ, or
cell not affected by the disorder or not intended to be targeted by
the therapeutic compound.
[0008] In some embodiments, the PL-inducing compound and the
therapeutic compound are administered at the same time or about the
same time before PL is induced in the patient. In some embodiments,
the therapeutic compound is administered only after PL is induced
in the patient. In some embodiments, the patient is treated with
the therapeutic compound for a treatment period and PL is induced
for a portion or over the entire duration of the treatment
period.
[0009] The method can further include monitoring the occurrence,
progress, or reversibility of PL in the patient. In some
embodiments, the monitoring step includes detecting the levels of
one or more biomarkers in a biological sample obtained from the
patient, the one or more biomarkers being selected from the group
consisting of 2,2' di-22:6-BMP, 3,2' di-22:6-BMP, 2,3' di-22:6-BMP,
3,3' di-22:6-BMP, di-18:1-BMP, di-18:2-BMP, 18:1/18:2-BMP,
18:1/22:6-BMP, 18:2/22:6-BMP, di-22:6-PG, di-18:1-PG, di-18:2-PG,
18:1/18:2-PG, 18:1/22:6-PG, 18:2/22:6-PG, mono-22:6-BMP,
mono-18:1-BMP, and mono-18:2-BMP. The monitoring step can further
include detecting the levels of one or more additional species of
BMP, PG, or mono-BMP, or total BMP. In some embodiments, the
therapeutic compound is administered after an elevated level of the
biomarker, as compared to a control level, is detected in the
biological sample.
[0010] The therapeutic compound can be a biological drug, e.g., an
antibody, antibody drug conjugate (ADC), protein, peptide,
peptidomimetic, peptoid, RNA, DNA, siRNA, miRNA, or RNA aptamer. In
some embodiments, the therapeutic compound is a small molecule
drug.
[0011] In some embodiments, the PL-inducing compound is selected
from the group consisting of ambroxol (metabolite of bromhexine),
amikacin, amiodarone, amitriptyline, aripiprazole, atorvastatin,
azithromycin, bedaquiline (TMC207, R207910), bepotastine,
bromopheniramine, busulfan, chlorcyclizine, chloropromazine,
chlorpheniramine (chlorphenamine), chloroquine, citalopram,
clarithromycin, clindamycin, cloforex (prodrug of
chlrophentermine), clomipramine, clozapine, crizotinib, cyclizine,
dronedarone, SR33589, duloxetine, erythromycin, escitalopram,
everolimus, RAD001, fluoxetine, gentamicin, haloperidol,
homochlorcyclizine, hydroxychloroquine, hydroxyzine, imipramine (G
22355, melipramine), indormamin, iprindole (pramindole), ketotifen,
kevodopa (L-DOPA), kaprotiline, meclizine, memantine,
nortriptyline, noxiptiline (noxiptyline or dibenzoxine),
paroxetine, pentamidine, pheniramine, phentermine, posaconazole,
promethazine, quinacrine (mepacrine), rapamycin, rosuvastatin,
sapropterin (tetrahydrobiopterin), simvastatin, tamoxifen,
telithromycin, tobramycin, trifluperazine, trimeprazine
(alimenazine), trimethoprim, tunicamycin, vandetanib, verenicline,
zonisamide, poloxamers (pluronics, synperonics, and kolliphor),
total parenteral nutrition (TPN) solutions, steroid hormones, and
oxysterols. One or more PL-inducing compounds can be used to induce
PL in a subject.
[0012] The therapeutic compound can be any of the PL-inducing
compounds listed above or selected from the group consisting of
abogovomab, abciximab, abagovomab, abciximab, abrilumab, actoxumab,
adalimumab, adecatumumab, aducanumab, afelimomab, afutuzumab,
alacizumab pegol, aLD518, alemtuzumab, alirocumab, altumomab
pentetate, amatuximab, anatumomab mafenatox, anetumab ravtansine,
anifrolumab, anrukinzumab, apolizumab, arcitumomab, ascrinvacumab,
aselizumab, atezolizumab, atinumab, atlizumab (tocilizumab),
atorolimumab, bapineuzumab, basiliximab, bavituximab, bectumomab,
begelomab, belimumab, benralizumab, bertilimumab, besilesomab,
bevacizumab, bezlotoxumab, biciromab, bimagrumab, bimekizumab,
bivatuzumab mertansine, blinatumomab, blosozumab, bococizumab,
brentuximab vedotin, briakinumab, brodalumab, brolucizumab,
brontictuzumab, canakinumab, cantuzumab mertansine, cantuzumab
ravtansine, caplacizumab, capromab pendetide, carlumab,
catumaxomab, cBR96-doxorubicin immunoconjugate, cedelizumab,
certolizumab pegol, cetuximab, citatuzumab bogatox, cixutumumab,
clazakizumab, clenoliximab, clivatuzumab tetraxetan, codrituzumab,
coltuximab ravtansine, conatumumab, concizumab, cR6261, crenezumab,
dacetuzumab, daclizumab, dalotuzumab, dapirolizumab pegol,
daratumumab, dectrekumab, demcizumab, denintuzumab mafodotin,
denosumab, derlotuximab biotin, detumomab, dinutuximab,
diridavumab, dorlimomab aritox, drozitumab, duligotumab, dupilumab,
durvalumab, dusigitumab, ecromeximab, eculizumab, edobacomab,
edrecolomab, efalizumab, efungumab, eldelumab, elgemtumab,
elotuzumab, elsilimomab, emactuzumab, emibetuzumab, enavatuzumab,
enfortumab vedotin, enlimomab pegol, enoblituzumab, enokizumab,
enoticumab, ensituximab, epitumomab cituxetan, epratuzumab,
erlizumab, ertumaxomab, etaracizumab, etrolizumab, evinacumab,
evolocumab, exbivirumab, fanolesomab, faralimomab, farletuzumab,
fasinumab, FBTA05, felvizumab, fezakinumab, ficlatuzumab,
figitumumab, firivumab, flanvotumab, fletikumab, fontolizumab,
foralumab, foravirumab, fresolimumab, fulranumab, futuximab,
galiximab, ganitumab, gantenerumab, gavilimomab, gemtuzumab
ozogamicin, gevokizumab girentuximab, glembatumumab vedotin,
golimumab, gomiliximab, guselkumab, ibalizumab, ibritumomab
tiuxetan, icrucumab, idarucizumab, igovomab, IMAB362, imalumab,
imciromab, imgatuzumab, inclacumab, indatuximab ravtansine,
indusatumab vedotin, infliximab, inolimomab, inotuzumab ozogamicin,
intetumumab, ipilimumab, iratumumab, isatuximab, itolizumab,
ixekizumab, keliximab, labetuzumab, lambrolizumab, lampalizumab,
lebrikizumab, lemalesomab, lenzilumab, lerdelimumab, lexatumumab,
libivirumab, lifastuzumab vedotin, ligelizumab, lilotomab
satetraxetan, lintuzumab, lirilumab, lodelcizumab, lokivetmab,
lorvotuzumab mertansine, lucatumumab, lulizumab pegol, lumiliximab,
lumretuzumab, mapatumumab, margetuximab, maslimomab, matuzumab,
mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab,
mirvetuximab soravtansine, mitumomab, mogamulizumab, morolimumab,
motavizumab, moxetumomab pasudotox, muromonab-CD3, nacolomab
tafenatox, namilumab, naptumomab estafenatox, narnatumab,
natalizumab, nebacumab, necitumumab, nemolizumab, nerelimomab,
nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan,
obiltoxaximab, obinutuzumab, ocaratuzumab, ocrelizumab, odulimomab,
ofatumumab, olaratumab, olokizumab, omalizumab, onartuzumab,
ontuxizumab, opicinumab, oportuzumab monatox, oregovomab,
orticumab, otelixizumab, otlertuzumab, oxelumab, ozanezumab,
ozoralizumab, pagibaximab, palivizumab, panitumumab, pankomab,
panobacumab, parsatuzumab, pascolizumab, pasotuxizumab,
pateclizumab, patritumab, pembrolizumab, pemtumomab, perakizumab,
pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin,
pintumomab, placulumab, polatuzumab vedotin, ponezumab, priliximab,
pritoxaximab, pritumumab, PRO 140, quilizumab, racotumomab,
radretumab, rafivirumab, ralpancizumab, ramucirumab, ranibizumab,
raxibacumab, refanezumab, regavirumab, reslizumab, rilotumumab,
rinucumab, rituximab, robatumumab, roledumab, romosozumab,
rontalizumab, rovelizumab, ruplizumab, sacituzumab govitecan,
samalizumab, sarilumab, satumomab pendetide, secukinumab,
seribantumab, setoxaximab, sevirumab, SGN-CD19A, SGN-CD33A,
sibrotuzumab, sifalimumab, siltuximab, simtuzumab, siplizumab,
sirukumab, sofituzumab vedotin, solanezumab, solitomab,
sonepcizumab, sontuzumab, stamulumab, sulesomab, suvizumab,
tabalumab, tacatuzumab tetraxetan, tadocizumab, talizumab,
tanezumab, taplitumomab paptox, tarextumab, tefibazumab, telimomab
aritox, tenatumomab, teneliximab, teplizumab, teprotumumab,
tesidolumab, tetulomab, TGN1412, ticilimumab (tremelimumab),
tigatuzumab, tildrakizumab, TNX-650, tocilizumab (atlizumab),
toralizumab, tosatoxumab, tositumomab, tovetumab, tralokinumab,
trastuzumab, trastuzumab emtansine, TRBS07, tregalizumab,
tremelimumab, trevogrumab, tucotuzumab, celmoleukin, tuvirumab,
ublituximab, ulocuplumab, urelumab, urtoxazumab, ustekinumab,
vandortuzumab vedotin, vantictumab, vanucizumab, vapaliximab,
varlilumab, vatelizumab, vedolizumab, veltuzumab, vepalimomab,
vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin,
votumumab, zalutumumab, zanolimumab, zatuximab, ziralimumab,
zolimomab aritox, etanercept, insulin glargine, pegfilgrastim,
salmon calcitonin, cyclosporine, octreotide, liraglutide,
bivalirudin, desmoprossin, interferon beta-1a, interferon beta-1a,
patisiran (ALN-TTR02), revusiran (ALN-TTRsc), fitusiran (ALN-AT3),
ALN-CCr, ALN-AS1, ALN-PCSsc, ALN-VSP02, siRNA-EphA2-DOPC, Atu027,
TKM-0880301, TKM-100201, ALN-RSV01, PRO-040201, ALN-TTR02,
CALAA-01, TD101, AGN211745, QPI-1007, I5NP, PF-655 (PF-04523655),
siG12D LODER, bevasiranib, SYL1001, SYL040012, SYL040012, CEQ508,
RXi-109, ARC-520, mipomersen, trabedersen (AP12009), fomivirsen,
iomitapide, brentuximab (Adcetris), and ado-trastuzamab emtanisine
(Kadcyla).
[0013] In another aspect, described herein is a pharmaceutical
composition containing a PL-inducing compound and a therapeutic
compound for a disorder. The PL-inducing compound and the
therapeutic compounds can be selected from the compounds listed
above.
[0014] In some embodiments, the composition is a
controlled-released formulation (e.g. delayed- or extended-release
formulation). In some embodiment, the formulation is formulated to
release the PL-inducing compound and the therapeutic compound
simultaneously or at different rates or times. In some embodiments,
the PL-inducing compound is released before the therapeutic
compound. The composition can be a target-released formulation. In
some embodiments, the composition is an implant or transdermal
device.
[0015] The details of one or more embodiments of the invention are
set forth in the accompanying drawings and the description below.
Other features, objects, and advantages of the invention will be
apparent from the description and drawings, and from the
claims.
DESCRIPTION OF DRAWINGS
[0016] FIG. 1 is a diagram showing structures of
bis(monoacylglycerol) phosphate (BMP) isoforms.
[0017] FIG. 2A is a diagram showing an exemplary therapeutic PL
regimen to inhibit the lysosomal degradation of biological drugs in
disease cells or tissues to improve in vivo efficacy.
[0018] FIG. 2B is a diagram showing an exemplary therapeutic PL
regimen to decrease the release of active toxins or drugs from ADCs
in healthy/non-targeted cells or tissues to improve in vivo
efficacy.
[0019] FIG. 3 is a set of electron micrographs depicting the
accumulation of lysosomal myeloid bodies, indicative of PL, in rat
tissues. Sprague-Dawley rats received once daily oral (PO)
co-administration of bedaquiline (60 mg/kg/day) and citalopram (70
mg/kg/day) for 2 weeks.
[0020] FIG. 4 is a set of graphs showing drug concentrations in
tissues of rats with PL induced by co-administration of bedaquiline
(60 mg/kg/day) and citalopram (70 mg/kg/day) for 2-weeks.
[0021] FIG. 5 is a set of graphs showing di-22:6-BMP concentrations
(including all isoforms) in tissues and urine of rats with PL
induced by co-administration of bedaquiline (60 mg/kg/day) and
citalopram (70 mg/kg/day) for 2-weeks.
[0022] FIG. 6 is a set of graphs showing that drug-induced PL
decreased lysosomal degradation of 14-3-3 proteins. Levels of
14-3-3 gamma proteins were higher in the livers of rats with
drug-induced PL compared to vehicle/controls. Urine di-22:6 BMP was
used to monitor PL.
[0023] FIG. 7 is a set of graphs showing that PL decreased the
activity of cathepsin-B, a lysosomal protease in rat tissues.
Samples that contain cathepsin-B cleaved the synthetic substrate
RR-AFC to release free AFC.
[0024] FIG. 8 is a set of graphs showing that PL-induction
increased drug tissue concentrations to enhance in vivo drug
efficacy.
[0025] FIG. 9 is a graph showing di-22:6-BMP concentrations
(including all isoforms) in tissues and urine of rats treated with
amiodarone alone (150 mg/kg/day) and co-administered with
fluoxetine (10 mg/kg/day) for 2-weeks.
[0026] FIG. 10 is a set of pie charts showing that PL increased the
efficacy of a drug targeted to a particular tissue by increasing
drug exposure in that tissue. Sprague-Dawley rats were treated with
amiodarone alone (150 mg/kg/day) or co-administered with fluoxetine
(10 mg/kg/day) for 2 weeks. Administration with fluoxetine shifted
amiodarone toward the target organ (heart) to increase efficacy and
away from lung to reduce amiodarone-induced pulmonary toxicity.
DETAILED DESCRIPTION
[0027] This disclosure is based, at least in part, on the
unexpected discovery that PL can be used to preferentially inhibit
lysosomal degradation and increase drug exposure in target tissues,
organs or cells and thereby enhance therapeutic efficacy.
Phospholipidosis
[0028] PL is induced by various natural and synthetic agents,
including widely used marketed drugs. It is characterized by the
excessive accumulation of multi-lamellar (myeloid) and/or zebra
bodies in the lysosomes of affected cells. Some tissues, such as
lung, kidney, and skin, normally contain myeloid bodies which
represent main storage sites for undigested and secreted materials.
See e.g., Schmitz and Muller, Journal of Lipid Research (1991)
32:1539-1570. Many cell and tissue types are reported as being
susceptible to drug-induced PL (e.g., liver, lung, kidney, and
heart). See e.g., Hruban, Environmental Health Perspectives (1984)
55:53-76. In this context, multi-lamellar bodies over accumulate in
various tissues and serve as repositories for drugs, drug
metabolites, undigested drug-phospholipid complexes, and other
undigested cellular materials. The pattern of PL is drug, species,
and tissue dependent. It is typically reversible after the
discontinuation of drug treatment. At a manageable level, PL can be
manifested in the absence of or without serious drug side effects.
See e.g., Cartwright et al., Toxicologic Pathology (2009)
37:902-910.
Biomarkers for Phospholipidosis
[0029] Myeloid bodies are rich in bis(monoacylglycerol)phosphate
(BMP), a phospholipid uniquely found in lysosomes and late
endosomes. BMP can theoretically exist in four geometrical
isoforms. See FIG. 1. Molecular species of BMP with different fatty
acid chains (e.g. 22:6, 18:1, and 18:2 fatty acids) correlate
differentially with the patterns of tissue PL induced by various
drugs. See US2010/0267061. An increase in BMP occurs in PL because
of its important roles in late endosomal/lysosomal degradation
pathways. BMP isoforms are sensitive and specific biomarkers of
tissue PL. They can be monitored in the plasma, serum, and urine.
BMP isoforms that can be used to monitor PL include 2,2'
di-22:6-BMP, 3,2' di-22:6-BMP, 2,3' di-22:6-BMP, 3,3' di-22:6-BMP,
di-18:1-BMP, di-18:2-BMP, 18:1/18:2-BMP, 18:1/22:6-BMP,
18:2/22:6-BMP, di-22:6-PG, di-18:1-PG, di-18:2-PG, 18:1/18:2-PG,
18:1/22:6-PG, 18:2/22:6-PG, mono-22:6-BMP, mono-18:1-BMP, and
mono-18:2-BMP. Techniques for detecting and quantifying BMP
isoforms are known in the art. See, e.g., US2010/0267061.
Lysosomal Degradation of Bio-Therapeutic Drugs
[0030] Lysosomes are responsible for the breakdown of
macromolecules (e.g. lipids, proteins, carbohydrates, DNA, and RNA)
derived from the extracellular space through endocytosis or
phagocytosis, and from the cytoplasm through autophagy. See FIG.
2A. Lysosomes contain resident hydrolases that permit the
collective degradation of all types of macromolecules, including
biologics (e.g. antibodies, proteins, peptides, and nucleic acids).
The acidic environment of the lysosomal lumen (pH 4.5-5.0)
facilitates the degradation process by loosening the structures of
macromolecules and is optimal for the activities of lysosomal
hydrolases. See e.g., Appleqvist et al., Annals of Clinical and
Laboratory Science (2012) 42(3):231-242; and Zhang et al., Acta
Biochim Biophys Sin (2009) 41: 437-445.
Lysosomal Release of Toxins from Antibody-Drug Conjugates
[0031] An antibody-drug conjugate (ADC) consists of an antibody
conjugated to a toxin or drug via a cleavable linker. ADCs are
designed to bind specific proteins (e.g., receptors) expressed on
the surface of cells (e.g. cancer cells) intended for treatment.
After entering the cell, the linker is cleaved within the lysosome,
thereby releasing the drug or toxin within the targeted cell. See
FIG. 2B. The toxin or drug remains inactive while conjugated to the
antibody.
[0032] For example, a peptide linker can be cleaved by lysosomal
proteases such as cathepsin B. Alternatively, when the ADC is in
the target cell, the antibody is degraded and the toxin is released
within the lysosome.
Phospholipidosis for Enhancing Drug Efficacy
[0033] Described herein are methods for enhancing the efficacy of
therapeutic compounds (e.g., biological drugs and traditional small
molecule drugs) by inducing PL. PL can inhibit the lysosomal
degradation of drugs or the release of drugs from ADCs, thereby
enhancing therapeutic efficacy in targeted cells, tissue or organ,
or protect non-targeted cells, tissues, or organ from exposure to
the drugs. See FIGS. 2A and 2B. PL can also be used to
preferentially concentrate traditional small molecule drugs in
target cells, tissues, or organs, and spare exposure to
non-targeted cells/tissues.
[0034] The PL-inducing compound used in the treatment methods can
be one that induces PL in a specific tissue, organ and/or cell type
intended to be targeted by the therapeutic compounds.
Alternatively, it can be one that induces PL systemically or in
multiple tissues, organs or cell types. PL can be induced by one or
a combination of compounds to optimize the PL conditions to
optimally inhibit lysosomal enzymatic activity for a particular
therapeutic compound.
[0035] To practice the treatment methods described herein, the
above-described BMP biomarkers can be used to monitor the
occurrence, progress, and reversibility of PL in humans or test
animals. A subject has PL if the level(s) of the biomarker(s) in a
biological sample obtained from the subject are at or above their
corresponding control levels. A control level can be the level
found in a biological sample from a control subject with PL (e.g.,
induced by a compound) or biological samples from a control group
of subjects with PL. In some instances, the control level can be
the level in a biological sample obtained from the subject to be
treated with the therapeutic compound before PL is induced in the
subject.
[0036] The biological sample can be a bodily fluid sample,
including but not limited to whole blood, plasma, serum, urine, and
saliva. It can also be a cell, cell fraction, or a cell culture.
The sample can also be a whole tissue, tissue slice, or tissue
fraction. The sample can also be isolated endocytic vesicles, such
as endosomes, lysosomes, and exosomes derived from cells and
tissues.
[0037] The methods can be used to improve the efficacy of drugs
used to treat various diseases, disorders, conditions and syndromes
such as cancer (e.g. lung cancer, breast cancer, prostate cancer,
colon cancer) and neurological diseases (e g Alzheimer's disease,
Parkinson's disease).
I. Drugs Subjected to Lysosomal Degradation
[0038] A number of drugs, such as antibodies, proteins, peptides,
and nucleic acid drugs, are internalized into cells by endocytosis
and then degraded by lysosomal enzymes. See FIG. 2A.
[0039] PL-inducing compounds can directly and/or indirectly (i.e.,
through the accumulation of undigested materials or change in
lysosomal pH) inhibit lysosomal enzyme activities. The induction of
PL by such compounds can be used to prevent the degradation of
therapeutic drugs (e.g. antibodies, proteins, peptides,
peptidomimetics, peptoids, RNA, DNA, siRNA, miRNA, or RNA aptamer)
by lysosomal hydrolases thereby increasing their in vivo efficacy.
See FIG. 2A. In particular, PL can be induced in diseased or target
cells, tissues, or organs to more specifically inhibit lysosomal
degradation of the drugs in those cells, tissues, or organs.
[0040] PL has been shown to inhibit lysosomal proteases such as
cathepsin B. For example, the PL-inducing chloroquine inhibited
cathepsin B1 activities. See, e.g., Wibo and Poole, The Journal of
Cell Biology (1974) 63:430-440. As described below, PL induced by
the antibiotic bedaquiline unexpectedly slowed the lysosomal
degradation of proteins in rat liver. See FIG. 6. PL induced by
amiodarone slowed the activity of cathepsin B in rat liver and
spleen. See FIG. 7.
[0041] To practice the treatment method, PL can be induced in a
subject to be treated with a therapeutic compound that is subjected
to lysosomal degradation. PL can be induced at the same time or
after the therapeutic compound is administered. The biomarkers
described herein can be used to monitor PL in the subject. For
example, the biomarkers can be used to determine when the
therapeutic drug should be administered. Once PL is induced, it is
maintained or intensified to optimize therapy until the desired
therapeutic effect is achieved or the therapeutic treatment is
completed. PL is discontinued or reversed (e.g., by discontinuing
the PL treatment) after completion of the therapeutic
treatment.
II. Drugs Activated by Lysosomal Processing
[0042] A number of drugs, such as ADCs, are internalized into cells
by endocytosis and activated through lysosomal processing. See FIG.
2B. A PL-inducing compound can be administered with an ADC
cleavable by a lysosomal protease to inhibit release of the drug in
a non-target organ, cell, or tissue. The drug is thereby released
preferentially within the target organ, cell, or tissue to enhance
drug efficacy. See FIG. 2B.
[0043] More specifically, a patient in need of an ADC is treated
with the ADC and one or more PL-inducing compounds to induce PL and
inhibit lysosomal protease activities in the cell, organ or tissue
not intended to be targeted by the ADC. PL can be induced at the
same time or after the ADC is administered. The biomarkers
described herein can be used to monitor PL in the subject and
determine when the ADC should be administered. For example, using
the method, healthy cells can be protected from an ADC intended to
target and kill cancer cells.
[0044] PL is maintained until the therapeutic effect or an
effective concentration of the ADC has been achieved in the target
cell, tissue, or organ. The PL-inducing treatment is then
discontinued after completion of the therapeutic treatment.
III. Drugs Targeted to Tissues, Organs or Cells by
Phospholipidosis
[0045] Therapeutic efficacy can also be enhanced by using PL to
increase the concentrations of small molecule drugs in target
cells, tissues or organs and to preferentially distribute them,
thereby increasing drug exposure in the target cells, tissues or
organs.
[0046] Thus, a patient in need of a therapeutic treatment can be
administered with a small molecule drug together with one or more
PL-inducing compounds that induce PL in the intended target cell,
tissue or organ of the drug. The drug is circulated and
concentrated within myeloid bodies and then released in a
controlled manner (by modulating PL using PL-inducing compounds as
monitored by biomarkers) within the target tissue, cell or organ
where the drug produces a therapeutic effect.
[0047] PL is maintained until the effect or an effective amount of
the therapeutic molecule has been achieved in the target cell,
tissue or organ. PL is then reversed after completion of the
therapeutic treatment.
[0048] For example, as described below, an increased concentration
of fluoxetine (Prozac) occurred in tissues, including the target
tissue brain, during co-administration with the PL-inducing
compound amiodarone as compared with fluoxetine alone. Fluoxetine
uptake in tissues increased with tissue di-22:6-BMP concentrations.
See FIG. 10
[0049] In another example, the method can be used to enhance the
efficacy of polymer drug conjugates and biologics that have been
modified to extend their in vivo half-lives (e.g., pegylated
proteins, pegylated peptides, and fusion proteins).
Pharmaceutical Compositions
[0050] To practice the above-described treatment methods, a
pharmaceutical composition containing one or more PL-inducing
compounds and a therapeutic drug can be used. The composition can
be formulated as a controlled-release (e.g., timed-release,
delayed-release, or extended-release) formulation.
[0051] The release profile of the formulation can be designed to
enhance the efficacy of a therapeutic drug as described above. For
example, different therapeutic regimens or controlled-release
formulations (e.g., different combinations of PL-inducing compounds
and therapeutic drugs, different release profiles, or different
orders/timings of administration) can be tested in test human
subjects or animal models.
[0052] For example, the formulation can be formulated to release a
PL-inducing compound and a therapeutic drug at the same time. In
another example, the formulation can be formulated to first release
a PL-inducing compound to induce PL in the patient and then release
a therapeutic drug. The release of either or both the PL-inducing
compound and the therapeutic drug can be sustained or stopped as
required to achieve the desired result. The PL biomarkers can be
used to determine and monitor the optimal dosing time for the
therapeutic compound. Methods and materials for designing and
producing controlled-release formulations are known in the art.
[0053] The specific examples below are to be construed as merely
illustrative, and not limitative of the remainder of the disclosure
in any way whatsoever. Without further elaboration, it is believed
that one skilled in the art can, based on the description herein,
utilize the present invention to its fullest extent. All
publications cited herein are hereby incorporated by reference in
their entirety. Further, any mechanism proposed below does not in
any way restrict the scope of the claimed invention.
Example 1
[0054] Sprague-Dawley rats were co-administered bedaquiline (60
mg/kg/day) and citalopram (70 mg/kg/day) in a 2-week repeat dose
study. Tissue sections (liver, kidney, and heart) were collected at
2 weeks for electron microscopic examination and PL
biomarker/proteomic evaluation.
[0055] Representative electron micrographs are shown in FIG. 3. A
treatment-dependent increase in multi-lamellar lysosomal inclusions
and electron dense deposits was observed in liver, kidney, and
heart, indicative of drug-induced PL. Concentrations of bedaquiline
and citalopram in tissues are shown in FIG. 4. A corresponding
increase in the biomarker di-22:6-BMP was observed with drug
concentrations and PL in rat tissues and urine. See FIG. 5.
[0056] In rats with PL, the levels of 14-3-3 gamma protein were
unexpectedly higher in liver compared to the vehicle/control group.
See FIG. 6. 14-3-3 proteins are cytosolic proteins involved in
regulating various intracellular signaling, cell cycling,
apoptosis, and transcription regulation processes. See, e.g.,
Tzivion et al., Oncogene (2001) 20:6331-6338. 14-3-3 proteins are
reported substrates of cathepsins (D, L, S, and B), a family of
lysosomal proteases. See, e.g., Appelqvist et al., Annals of
Clinical and Laboratory Science (2012) 42(3):231-242; and Zavr nik
et al., Biochemical and Biophysical Research Communications (2015)
465(2):213-217.
[0057] It is proposed that the unexpected increase in liver 14-3-3
gamma was due to PL inhibition of lysosomal protease activity
either within the lysosomal or release to the cytosol. Urine
di-22:6-BMP isoforms can be used to monitor drug concentrations to
induce PL in specific tissues and modulate lysosomal enzyme
activity.
Example 2
[0058] Cathepsin-B is a lysosomal protease that plays an important
role in intracellular proteolysis. Cathepsin-B activity was
investigated in tissues from a 2-week repeat dose study of
amiodarone (150 mg/kg/day) in Sprague-Dawley rats. Liver and spleen
homogenates were analyzed using a fluorescence-based assay that
utilized the preferred cathepsin-B substrate sequence RR labeled
with amino-4-trifluoromethyl coumarin (AFC). Samples that contain
cathepsin-B cleave the synthetic substrate RR-AFC to release free
AFC.
[0059] The results are shown in FIG. 7. Unexpectedly, the levels of
free AFC were lower in tissues with amiodarone-induced PL compared
to controls, indicative of an inhibitory effect of PL on
cathepsin-B activity.
Example 3
[0060] Sprague-Dawley rats were treated with fluoxetine (10
mg/kg/day) alone or co-administered with amiodarone (150 mg/kg/day)
for 2 weeks. Tissues were collected at necropsy at 2 weeks. PL was
induced as indicated by an increase in the di-22:6-BMP biomarker in
various tissues. See FIG. 8.
[0061] Unexpectedly, fluoxetine concentrations were increased in
tissues with amiodarone-induced PL compared to administration of
fluoxetine alone. See FIG. 8. The results demonstrate PL can be
used to enhance drug tissue uptake and thereby may enhance drug
efficacy.
Example 4
[0062] Sprague-Dawley rats were treated with amiodarone alone (150
mg/kg/day) or co-administered with fluoxetine (10 mg/kg/day) for 2
weeks. Heart, liver, kidney and lung tissues were collected at
necropsy at 2 weeks. PL was induced in tissues of both treatment
groups as indicated by an increase in the PL biomarker di-22:6-BMP.
See FIG. 9.
Example 5
[0063] The time to onset of action of amiodarone is often long in
patients treated for arrhythmias. One reason might be a slow entry
of the drug into the target organ, the heart. See, e.g. Barbieri et
al., J Am Coll Cardiol (1986) July; 8(1):210-3. Co-administration
of amiodarone with fluoxetine in rats (as described above)
unexpectedly changed the distribution of amiodarone in rat tissues
and increased the amiodarone concentration in the heart. See FIG.
10.
[0064] Additionally, amiodarone-induced pulmonary toxicity is a
known serious complication of amiodarone therapy. See, e.g.,
Wolkove et al., Canadian Respiratory Journal (2009) 16(2):43-48.
The PL-inducer therapy decreased the concentration of amiodarone in
the lung tissue, thereby reducing the risk of pulmonary
toxicity.
OTHER EMBODIMENTS
[0065] All of the features disclosed in this specification may be
combined in any combination. Each feature disclosed in this
specification may be replaced by an alternative feature serving the
same, equivalent, or similar purpose. Thus, unless expressly stated
otherwise, each feature disclosed is only an example of a generic
series of equivalent or similar features.
[0066] A number of embodiments of the invention have been
described. Nevertheless, it will be understood that various
modifications may be made without departing from the spirit and
scope of the invention. Accordingly, other embodiments are within
the scope of the following claims.
* * * * *